Expression cassettes for regulation of expression in monocotyledonous plants

Abstract

The present invention relates to expression cassettes comprising at least one transcription regulating nucleotide sequence obtainable from the group of genes of monocotyle-donous plants consisting of caffeoyl-CoA-O-methyltransferase genes, C8,7-sterol isomerase genes, hydroxyproline-rich glycoprotein (HRGP) genes, lactate dehydrogenase genes, and chloroplast protein 12 like genes. More preferably the transcription regulating sequences are obtainable from Zea mays or Oryza sativa . The transcription regulating sequences are especially useful for root/kernel-preferential, leaf/endosperm-preferential, root/silk/kernel-preferential, or constitutive expression.

The invention claimed is: 
     
       1. An expression cassette for regulating expression in a monocotyledonous plant, said expression cassette comprising:
 i) a promoter nucleotide sequence sharing at least 98% sequence identity to the nucleotide sequence of SEQ ID NO:1; 
 ii) at least one nucleic acid sequence which is heterologous in relation to said promoter; and 
 iii) a terminator comprising a nucleotide sequence sharing at least 98% sequence identity to the nucleotide sequence of SEQ ID NO:34; 
 wherein the expression of the at least one nucleic acid sequence is constitutive. 
 
     
     
       2. The expression cassette of  claim 1 , further comprising the 5′-untranslated region of a plant expressed gene, or an intron from a plant expressed gene. 
     
     
       3. The expression cassette of  claim 2 , wherein the intron is from ubiquitin, actin, or alcohol dehydrogenase gene. 
     
     
       4. The expression cassette of  claim 1 , wherein the terminator comprises the nucleotide sequence of SEQ ID NO: 34. 
     
     
       5. A vector comprising the expression cassette of  claim 1 . 
     
     
       6. A transgenic host cell or non-human organism comprising:
 i) the expression cassette of  claim 1 ; or 
 ii) a vector comprising the expression cassette. 
 
     
     
       7. A transgenic plant comprising:
 i) the expression cassette of  claim 1 ; or 
 ii) a vector comprising the expression cassette. 
 
     
     
       8. A method for providing a transgenic expression cassette for heterologous expression in a monocotyledonous plant, said method comprising:
 I. isolating a promoter nucleotide sequence from a monocotyledonous plant, wherein the promoter nucleotide sequence shares at least 98% sequence identity to the nucleotide sequence of SEQ ID NO:1; 
 II. functionally linking said promoter nucleotide sequence to another nucleotide sequence of interest, which is heterologous in relation to said promoter nucleotide sequence; and 
 III. functionally linking said another nucleotide sequence of interest to a terminator comprising a nucleotide sequence sharing at least 98% sequence identity to the nucleotide sequence of SEQ ID NO:34. 
 
     
     
       9. The method of  claim 8 , wherein said terminator comprises the nucleotide sequence of SEQ ID NO: 34. 
     
     
       10. A cell culture, part, organ, tissue or transgenic propagation material derived from the non-human organisms of  claim 6 , wherein said cell culture, part, organ, tissue or transgenic propagation material comprises the expression cassette. 
     
     
       11. A method for the transgenic expression of a nucleic acid, said method comprising growing or culturing the non-human organisms of  claim 6  or cell cultures, parts, organs, tissues or transgenic propagation material derived therefrom, wherein said cell cultures, parts, organs, tissues or transgenic propagation material comprise the expression cassette. 
     
     
       12. A method for identifying and/or isolating a sequence with constitutive expression activity, said method comprising:
 a) obtaining a polynucleotide sequence sharing at least 98% sequence identity with the nucleic acid sequence of SEQ ID NO:1; 
 b) preparing an expression cassette comprising the polynucleotide operably linked to a reporter gene or marker and to a terminator sharing at least 98% sequence identity with the nucleic acid sequence of SEQ ID NO: 34; 
 c) testing the expression cassette for constitutive expression; and 
 d) identifying and/or isolating the polynucleotide with constitutive expression activity. 
 
     
     
       13. A method for directing expression in a monocotyledonous plant, said method comprising:
 I. introducing into a plant cell the expression cassette of  claim 1 , 
 II. selecting a transgenic cell which comprises said expression cassette, and 
 III. regenerating a plant from the transgenic cell, wherein the promoter nucleotide sequence directs constitutive expression of the nucleic acid sequence which is heterologous in relation to said promoter in the plant.